In Situ Studies of Mitochondrial Translation by Cryo-Electron Tomography

Robert Englmeier; Friedrich Förster

doi:10.1007/978-1-0716-0834-0_18

In Situ Studies of Mitochondrial Translation by Cryo-Electron Tomography

Robert Englmeier, Friedrich Förster^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic

Abstract

Cryo-electron tomography (cryo-ET) enables the three-dimensional (3D) visualization of macromolecular complexes in their native environment (in situ). The ability to visualize macromolecules in situ is in particular advantageous for complex, membrane-associated processes, such as mitochondrial translation. Mitochondrial translation occurs almost exclusively associated with the inner mitochondrial membrane, giving rise to the mitochondrial DNA-encoded subunits of oxidative phosphorylation machinery. In cryo-ET, the 3D volume is reconstructed from a set of 2D projections of a frozen-hydrated specimen, which is sequentially tilted and imaged at different angles in a transmission electron microscope. In combination with subtomogram analysis, cryo-ET enables the structure determination of macromolecular complexes and their 3D organization. In this chapter, we summarize all steps required for structural characterization of mitochondrial ribosomes in situ, ranging from data acquisition to tomogram reconstruction and subtomogram analysis.

Original language	English
Pages (from-to)	243-268
Number of pages	26
Journal	Methods in Molecular Biology
Volume	2192
DOIs	https://doi.org/10.1007/978-1-0716-0834-0_18
Publication status	Published - 2021

Bibliographical note

Funding Information:
We would like to thank Dr Stuart Howes for critical comments on the manuscript. This work was supported by funding from the Nederlandse Organisatie voor Wetenschappelijke Onderzoek (Vici 724.016.001), the European Research Council under the European Union?s Horizon2020 Programme (ERC Consolidator Grant Agreement 724425?BENDER) and the Deutsche Forschungsgemeinschaft (FO 716/3-2).

Publisher Copyright:
© 2021, Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

Cryo-electron tomography
Mitochondria
Mitochondrial ribosomes
Translation

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978-1-0716-0834-0_18Final published version, 880 KBLicence: Taverne

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abstract = "Cryo-electron tomography (cryo-ET) enables the three-dimensional (3D) visualization of macromolecular complexes in their native environment (in situ). The ability to visualize macromolecules in situ is in particular advantageous for complex, membrane-associated processes, such as mitochondrial translation. Mitochondrial translation occurs almost exclusively associated with the inner mitochondrial membrane, giving rise to the mitochondrial DNA-encoded subunits of oxidative phosphorylation machinery. In cryo-ET, the 3D volume is reconstructed from a set of 2D projections of a frozen-hydrated specimen, which is sequentially tilted and imaged at different angles in a transmission electron microscope. In combination with subtomogram analysis, cryo-ET enables the structure determination of macromolecular complexes and their 3D organization. In this chapter, we summarize all steps required for structural characterization of mitochondrial ribosomes in situ, ranging from data acquisition to tomogram reconstruction and subtomogram analysis.",

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author = "Robert Englmeier and Friedrich F{\"o}rster",

note = "Funding Information: We would like to thank Dr Stuart Howes for critical comments on the manuscript. This work was supported by funding from the Nederlandse Organisatie voor Wetenschappelijke Onderzoek (Vici 724.016.001), the European Research Council under the European Union?s Horizon2020 Programme (ERC Consolidator Grant Agreement 724425?BENDER) and the Deutsche Forschungsgemeinschaft (FO 716/3-2). Publisher Copyright: {\textcopyright} 2021, Springer Science+Business Media, LLC, part of Springer Nature.",

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doi = "10.1007/978-1-0716-0834-0_18",

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In Situ Studies of Mitochondrial Translation by Cryo-Electron Tomography

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Keywords

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